Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

Tanahashi, Masakazu; Kaneko, Ryuji; Hirata, Yukari; Hamajima, Makoto; Arinobu, Tetsuya; Ogawa, Tadashi; Ishii, Akira

doi:10.1007/s11419-010-0098-6

Cited by 24 publications

(9 citation statements)

References 20 publications

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“…Several methods are available in the literature for the analysis and quantification of ␣-amanitin in different matrices, mainly mushrooms [11][12][13][14][15], plasma [20,28,29] and urine [16,[21][22]29] Fig. 4.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, there are several high-performance liquid chromatography (HPLC)-ultraviolet (UV) [11][12][13][14][15][16][17], liquid chromatography (LC)-mass spectrometry (MS) [18][19][20] and ultra performance liquid chromatography (UPLC)-MS/MS [21] methods to determine ␣-amanitin in mushrooms, liver and biological fluids (Table 1). HPLC-MS methods for ␣-amanitin detection are accurate, sensitive, and specific; nevertheless HPLC-MS is an expensive technique and is not available in many laboratories.…”

Section: Introductionmentioning

confidence: 99%

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Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Garcia

Costa

Baptista

et al. 2015

Journal of Chromatography B

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α-Amanitin is a natural bicyclic octapeptide, from the family of amatoxins, present in the deadly mushroom species Amanita phalloides. The toxicological and clinical interests raised by this toxin, require highly sensitive, accurate and reproducible quantification methods for pharmacokinetic studies. In the present work, a high-performance liquid chromatographic (HPLC) method with in-line connected diode-array (DAD) and electrochemical (EC) detection was developed and validated to quantify α-amanitin in biological samples (namely liver and kidney). Sample pre-treatment consisted of a simple and unique deproteinization step with 5% perchloric acid followed by centrifugation at 16,000×g, 4°C, for 20min. The high recovery found for α-amanitin (≥96.8%) makes this procedure suitable for extracting α-amanitin from liver and kidney homogenates. The resulting supernatant was collected and injected into the HPLC. Mobile phase was composed by 20% methanol in 50mM citric acid, and 0.46mM octanessulfonic acid, adjusted to pH 5.5. The chromatographic runs took less than 22min and no significant endogenous interferences were observed at the α-amanitin retention time. Calibration curves were linear with regression coefficients higher than 0.994. The overall inter- and intra-assay precision did not exceed 15.3%. The present method has low interferences with simple and fast processing steps, being a suitable procedure to support in vivo toxicokinetic studies involving α-amanitin. In fact, the validated method was successfully applied to quantify α-amanitin in biological samples following intraperitoneal α-amanitin administration to rats. Moreover, human plasma was also used as matrix and the purposed method was adequate for detection of α-amanitin in that matrix. The results clearly indicate that the proposed method is suitable to investigate the pharmacokinetic and tissue distribution of α-amanitin. Additionally, the method will be very useful in the development of novel and potent antidotes against amatoxins poisoning and to improve the knowledge of α-amanitin toxicity.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Garcia

Costa

Baptista

et al. 2015

Journal of Chromatography B

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“…A lack of analytical methods to quantify α-amanitin in biological samples hinders the quantification of this toxin in tissues. According to the literature, liquid chromatography is the method of excellence for α-amanitin quantification (131,132).…”

Section: Polymyxin B Prevented α-Amanitin-induced Renal and Hepatic Dmentioning

confidence: 99%

Discovery of an effective antidote for Amanita phalloides poisoning

et al. 2015

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“…In the three reports [11,36,37] on LC-MS analysis of amanitins, octadecyl columns were used. However, separation of amanitins was not satisfactory.…”

Section: Separation Columnmentioning

confidence: 99%

“…Although the results by LC-TOFMS were satisfactory, those obtained by the LC-tandem MS gave too many peaks of product ions, which were much less intense than each precursor ion (protonated molecular ion). Tanahashi et al[11] also tried the usual LC-MS-MS analysis for a-amanitin and b-amanitin in human plasma; in the MS-MS mode, a-amanitin and b-amanitin gave base peaks at m/z 901 and 902, because of [M-18?H] ? , respectively, but the intensities of the product ions were estimated to be only about one tenth…”

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confidence: 99%

Mushroom toxins: a forensic toxicological review

et al. 2011

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Mushrooms are ubiquitous in the world. Amateur hunters harvest mushrooms growing in forests to enjoy eating them as seasonal delicacies, and occasionally they cause poisonings and even deaths. In this review, mushroom toxins are tabulated according to mushroom species, symptoms, toxicities and analytical methods on the basis of references. Second, because we constructed a method for analysis of amatoxins, the most virulent mushroom toxins, by liquid chromatography-time-of-flight-mass spectrometry, we introduce it for use in forensic toxicology. Third, an extensive poisoning incident after consumption of the usually edible mushroom Pleurocybella porrigens took place in nine prefectures in Japan from September to December 2004, resulting in 59 poisoned people including 19 deaths; this incident is briefly described and discussed in relation to its causative toxin(s). Finally, we present the chemical structures of new toxins purified from the highly toxic mushrooms Podostroma cornu-damae and Russula subnigricans; their structures were very unique, and the toxicities were comparable to those of amatoxins. From the forensic toxicological point of view, reports on sophisticated methodology for analyses of mushroom toxins seem to be too scant even for the well-known toxins. Hereafter, a number of toxic mushrooms and their new toxins are expected to be disclosed, especially because of environmental changes such as the global warming phenomenon.

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Simple analysis of α-amanitin and β-amanitin in human plasma by liquid chromatography-mass spectrometry

Cited by 24 publications

References 20 publications

Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Quantification of alpha-amanitin in biological samples by HPLC using simultaneous UV- diode array and electrochemical detection

Discovery of an effective antidote for Amanita phalloides poisoning

Mushroom toxins: a forensic toxicological review

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